Lettuce variety chalone

ABSTRACT

The present invention provides novel lettuce cultivar Chalone and plant parts, seed, and tissue culture therefrom. The invention also provides methods for producing a lettuce plant by crossing the lettuce plants of the invention with themselves or another lettuce plant. The invention also provides lettuce plants produced from such a crossing as well as plant parts, seed, and tissue culture therefrom.

FIELD OF THE INVENTION

This invention is in the field of lettuce plants.

BACKGROUND OF THE INVENTION

The present invention relates to a lettuce (Lactuca sativa L.) varietydesignated Chalone.

Practically speaking, all cultivated forms of lettuce belong to thehighly polymorphic species Lactuca sativa that is grown for its ediblehead and leaves. Lactuca sativa is in the Cichoreae tribe of theAsteraceae (Compositae) family. Lettuce is related to chicory,sunflower, aster, dandelion, artichoke, and chrysanthemum. Sativa is oneof about 300 species in the genus Lactuca. There are seven differentmorphological types of lettuce. The crisphead group includes the icebergand batavian types. Iceberg lettuce has a large, firm head with a crisptexture and a white or creamy yellow interior. The batavian lettucepredates the iceberg type and has a smaller and less firm head. Thebutterhead group has a small, soft head with an almost oily texture. Theromaine, also known as cos lettuce, has elongated upright leaves forminga loose, loaf-shaped head and the outer leaves are usually dark green.Leaf lettuce comes in many varieties, none of which form a head, andinclude the green oak leaf variety. Latin lettuce looks like a crossbetween romaine and butterhead. Stem lettuce has long, narrow leaves andthick, edible stems. Oilseed lettuce is a type grown for its large seedsthat are pressed to obtain oil. Latin lettuce, stem lettuce, and oilseedlettuce are seldom seen in the United States.

Presently, there are over one thousand known lettuce cultivars. As acrop, lettuce is grown commercially wherever environmental conditionspermit the production of an economically viable yield.

Lettuce, in general, and leaf lettuce in particular, is an important andvaluable vegetable crop. Thus, there is an ongoing need for improvedlettuce varieties.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel lettuce cultivardesignated Chalone, also known as LICU18-0048, having desirablecharacteristics including heavy head weight and large headcircumference. The invention also encompasses the seeds of lettucecultivar Chalone, the plants of lettuce cultivar Chalone, plant parts ofthe lettuce cultivar Chalone (including leaves, seed, gametes), methodsof producing seed from lettuce cultivar Chalone, and method forproducing a lettuce plant by crossing the lettuce cultivar Chalone withitself or another lettuce plant, methods for producing a lettuce plantcontaining in its genetic material one or more transgenes, and thetransgenic lettuce plants produced by that method. The invention alsorelates to methods for producing other lettuce plants derived fromlettuce cultivar Chalone and to lettuce plants, parts thereof and seedderived by the use of those methods. The present invention furtherrelates to hybrid lettuce seeds and plants (and parts thereof includingleaves) produced by crossing lettuce cultivar Chalone with anotherlettuce plant.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of lettuce cultivar Chalone. In embodiments, thetissue culture is capable of regenerating plants having all oressentially all of the physiological and morphological characteristicsof the foregoing lettuce plant and/or of regenerating plants having thesame or substantially the same genotype as the foregoing lettuce plant.In exemplary embodiments, the regenerable cells in such tissue culturesare meristematic cells, cotyledons, hypocotyl, leaves, pollen, embryos,roots, root tips, anthers, pistils, ovules, shoots, stems, petiole,pith, flowers, capsules and/or seeds as well as callus and/orprotoplasts derived from any of the foregoing. Still further, thepresent invention provides lettuce plants regenerated from the tissuecultures of the invention.

As a further aspect, the invention provides a method of producinglettuce seed, the method comprising crossing a plant of lettuce cultivarChalone with itself or a second lettuce plant. Optionally, the methodfurther comprises collecting the seed.

Another aspect of the invention provides methods for producing hybridsand other lettuce plants derived from lettuce cultivar Chalone. Lettuceplants derived by the use of those methods are also part of theinvention as well as plant parts, seed, gametes and tissue culture fromsuch hybrid or derived lettuce plants.

In representative embodiments, a lettuce plant derived from lettucecultivar Chalone comprises cells comprising at least one set ofchromosomes derived from lettuce cultivar Chalone. In embodiments, alettuce plant or population of lettuce plants derived from lettucecultivar Chalone comprises, on average, at least 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of its alleles (i.e., theoretical allelic content; TAC) fromlettuce cultivar Chalone, e.g., at least about 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of the genetic complement of lettuce cultivar Chalone, andoptionally is the result of a breeding process comprising one or twobreeding crosses and one or more of selfing, sibbing, backcrossingand/or double haploid techniques in any combination and any order. Inembodiments, the breeding process does not include a breeding cross, andcomprises selfing, sibbing, backcrossing and or double haploidtechnology. In embodiments, the lettuce plant derived from lettucecultivar Chalone is one, two, three, four, five or more breeding crossesremoved from lettuce cultivar Chalone.

In embodiments, a hybrid or derived plant from lettuce cultivar Chalonecomprises a desired added trait(s). In representative embodiments, alettuce plant derived from lettuce cultivar Chalone comprises at least10 of the morphological and physiological characteristics of lettucecultivar Chalone (e.g., as described in Table 2 and Tables 1 and 3-16).In embodiments, the lettuce plant derived from lettuce cultivar Chalonecomprises essentially all of the morphological and physiologicalcharacteristics of lettuce cultivar Chalone (e.g., cultivar Chalonecomprises at least 10 of the morphological and physiologicalcharacteristics of lettuce cultivar Chalone (e.g., as described in Table2 and Tables 1 and 3-16).

The invention also relates to methods for producing a lettuce plantcomprising in its genetic material one or more transgenes and to thetransgenic lettuce plant produced by those methods (and progeny lettuceplants comprising the transgene). Also provided are plant parts, seedand tissue culture from such transgenic lettuce plants, optionallywherein one or more cells in the plant part, seed, or tissue culturecomprises the transgene. The transgene can be introduced via planttransformation and/or breeding techniques.

In another aspect, the present invention provides for single locusconverted plants of lettuce cultivar Chalone. Plant parts, seed, andtissue culture from such single locus converted plants are alsocontemplated by the present invention. The single locus may be adominant or recessive allele. In representative embodiments, the singletransferred locus confers such traits as male sterility, herbicideresistance, pest resistance (e.g., insect and/or nematode resistance),modified fatty acid metabolism, modified carbohydrate metabolism,disease resistance (e.g., for bacterial, fungal and/or viral disease),male fertility, enhanced nutritional quality, improved appearance (e.g.,color), improved salt tolerance, industrial usage, or any combinationthereof. The single locus may be a naturally occurring lettuce locus, agenome edited locus, a mutated locus (e.g., chemically or radiationinduced), or a transgene introduced into lettuce through geneticengineering techniques.

The invention further provides methods for developing lettuce plants ina lettuce plant breeding program using plant breeding techniquesincluding, for example, recurrent selection, backcrossing, pedigreebreeding, double haploid techniques, restriction fragment lengthpolymorphism enhanced selection, genetic marker enhanced selectionand/or transformation. Seeds, lettuce plants, and parts thereof,produced by such breeding methods are also part of the invention.

The invention also provides methods of multiplication or propagation oflettuce plants of the invention, which can be accomplished using anymethod known in the art, for example, via vegetative propagation and/orseed.

The invention further provides a method of producing food or feedcomprising (a) obtaining a lettuce plant of the invention, optionallywherein the plant has been cultivated to maturity, and (b) collecting atleast one lettuce plant or part thereof (e.g., leaves) from the plant.

Additional aspects of the invention include harvested products andprocessed products from the lettuce plants of the invention. A harvestedproduct can be a whole plant or any plant part, as described herein.Thus, in some embodiments, a non-limiting example of a harvested productincludes a seed, a leaf and/or a stem.

In representative embodiments, a processed product includes, but is notlimited to: cut, sliced, ground, pureed, dried, canned, jarred, washed,packaged, frozen and/or heated leaves and/or seeds of the lettuce plantsof the invention, or any other part thereof. In embodiments, theprocessed product includes washed and packaged leaves (or parts thereof)of the invention.

The seed of the invention can optionally be provided as an essentiallyhomogenous population of seed of a single plant or cultivar. Essentiallyhomogenous populations of seed are generally free from substantialnumbers of other seed, e.g., at least about 90%, 95%, 96%, 97%, 98% or99% pure.

In representative embodiments, the invention provides a seed of lettucecultivar Chalone.

As a further aspect, the invention provides a plant of lettuce cultivarChalone.

As an additional aspect, the invention provides a lettuce plant, or apart thereof, having all or essentially all of the physiological andmorphological characteristics of a plant of lettuce cultivar Chalone.

As another aspect, the invention provides leaves and/or seed of thelettuce plants of the invention and a processed product from the leavesand/or seed of the inventive lettuce plants.

As still another aspect, the invention provides a method of producinglettuce seed, the method comprising crossing a lettuce plant of theinvention with itself or a second lettuce plant. The invention alsoprovides seed produced by this method and plants produced by growing theseed.

As yet a further aspect, the invention provides a method for producing aseed of a lettuce plant derived from lettuce cultivar Chalone, themethod comprising: (a) crossing a lettuce plant of lettuce cultivarChalone with a second lettuce plant; and (b) allowing seed of a lettuceplant derived from lettuce cultivar Chalone to form. In embodiments, themethod further comprises: (c) growing a plant from the seed derived fromlettuce cultivar Chalone of step (b); (d) selfing the plant grown fromthe lettuce seed derived from lettuce cultivar Chalone or crossing it toa second lettuce plant to form additional lettuce seed derived fromlettuce cultivar Chalone, and (e) repeating steps (c) and (d) 0 or moretimes to generate further derived lettuce seed. Optionally, the methodcomprises: (e) repeating steps (c) and (d) one or more times (e.g., oneto three, one to five, one to six, one to seven, one to ten, three tofive, three to six, three to seven, three to eight or three to tentimes) to generate further derived lettuce plants. As another option,the method can comprise collecting the seed. The invention also providesseed produced by these methods and plants produced by growing the seed.

As another aspect, the invention provides a method of producing lettuceleaves, the method comprising: (a) obtaining a plant of lettuce cultivarChalone, optionally wherein the plant has been cultivated to maturity;and (b) collecting leaves from the plant. The invention also providesthe leaves produced by this method.

Still further, as another aspect, the invention provides a method ofvegetatively propagating a plant of lettuce cultivar Chalone. In anon-limiting example, the method comprises: (a) collecting tissuecapable of being propagated from a plant of lettuce cultivar Chalone;(b) cultivating the tissue to obtain proliferated shoots; and (c)rooting the proliferated shoots to obtain rooted plantlets. Optionally,the invention further comprises growing plants from the rootedplantlets. The invention also encompasses the plantlets and plantsproduced by these methods.

As an additional aspect, the invention provides a method of introducinga desired added trait into lettuce cultivar Chalone, the methodcomprising: (a) crossing a first plant of lettuce cultivar Chalone witha second lettuce plant that comprises a desired trait to produce F₁progeny; (b) selecting an F₁ progeny that comprises the desired trait;(c) crossing the selected F₁ progeny with lettuce cultivar Chalone toproduce backcross progeny; and (d) selecting backcross progenycomprising the desired trait to produce a plant derived from lettucecultivar Chalone comprising a desired trait. In embodiments, theselected progeny comprises all or essentially all the morphological andphysiological characteristics of the first plant of lettuce cultivarChalone. Optionally, the method further comprises: (e) repeating steps(c) and (d) one or more times in succession (e.g., one to three, one tofive, one to six, one to seven, one to ten, three to five, three to six,three to seven, three to eight or three to ten times) to produce a plantderived from lettuce cultivar Chalone comprising the desired trait.

In representative embodiments, the invention also provides a method ofproducing a plant of lettuce cultivar Chalone comprising a desired addedtrait, the method comprising introducing a transgene conferring thedesired trait into a plant of lettuce cultivar Chalone. The transgenecan be introduced by transformation methods (e.g., genetic engineering)or breeding techniques. In embodiments, the plant comprising thetransgene comprises all or essentially all of the morphological andphysiological characteristics of lettuce cultivar Chalone.

The invention also provides lettuce plants produced by the methods ofthe invention, wherein the lettuce plant has the desired added trait aswell as seed from such lettuce plants.

According to the foregoing methods, the desired added trait can be anysuitable trait known in the art including, for example, male sterility,male fertility, herbicide resistance, insect or pest (e.g., insectand/or nematode) resistance, modified fatty acid metabolism, modifiedcarbohydrate metabolism, disease resistance (e.g., for bacterial, fungaland/or viral disease), enhanced nutritional quality, increasedsweetness, increased flavor, improved ripening control, improved salttolerance, industrial usage, or any combination thereof.

In representative embodiments, a transgene conferring herbicideresistance confers resistance to glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine,benzonitrile, or any combination thereof.

In representative embodiments, a transgene conferring pest resistance(e.g., insect and/or nematode resistance) encodes a Bacillusthuringiensis endotoxin.

In representative embodiments, transgenic plants, single locus convertedplants, hybrid plants and lettuce plants derived from lettuce cultivarChalone have at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of themorphological and physiological characteristics of lettuce cultivarChalone (e.g., as described in Table 2 and Tables 1 and 3-16) in anycombination, or even all or essentially all of the morphological andphysiological characteristics of lettuce cultivar Chalone, so that saidplants are not significantly different for said traits than lettucecultivar Chalone, as determined at the 5% significance level when grownin the same environmental conditions; optionally, with the presence ofone or more desired additional traits (e.g., male sterility, diseaseresistance, pest or insect resistance, herbicide resistance, and thelike).

In embodiments, the plants of the invention have at least 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more of the morphological and physiologicalcharacteristics of lettuce cultivar Chalone (e.g., as described in Table2 and Tables 1 and 3-16).

The invention also encompasses plant parts, plant material, pollen,ovules, leaves, fruit and seed from the lettuce plants of the invention.Also provided is a tissue culture of regenerable cells from the lettuceplants of the invention, where optionally, the regenerable cells are:(a) embryos, meristem, leaves, pollen, cotyledons, hypocotyls, roots,root tips, anthers, flowers, pistils, ovules, seed, shoots, stems,stalks, petioles, pith and/or capsules; or (b) callus or protoplastsderived from the cells of (a). Further provided are lettuce plantsregenerated from a tissue culture of the invention.

In still yet another aspect, the invention provides a method ofdetermining a genetic characteristic of lettuce cultivar Chalone or aprogeny thereof, e.g., a method of determining a genotype of lettucecultivar Chalone or a progeny thereof using molecular genetictechniques. In embodiments, the method comprises detecting in the genomeof a Chalone plant, or a progeny plant thereof, at least a firstpolymorphism, e.g., comprises nucleic acid amplification and/or nucleicacid sequencing. To illustrate, in embodiments, the method comprisesobtaining a sample of nucleic acids from the plant and detecting atleast a first polymorphism in the nucleic acid sample (e.g., using oneor more molecular markers). Optionally, the method may comprisedetecting a plurality of polymorphisms (e.g., two or more, three ormore, four or more, five or more, six or more, eight or more or ten ormore polymorphisms, etc.) in the genome of the plant. In representativeembodiments, the method further comprises storing the results of thestep of detecting the polymorphism(s) on a computer readable medium. Theinvention further provides a computer readable medium produced by such amethod.

In addition to the exemplary aspects and embodiments described above,the invention is described in more detail in the description of theinvention set forth below.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the development of a novellettuce cultivar having desirable characteristics including heavy headweight and large head circumference.

It should be appreciated that the invention can be embodied in differentforms and should not be construed as limited to the embodiments setforth herein. Rather, these embodiments are provided so that thisdisclosure will be thorough and complete, and will fully convey thescope of the invention to those skilled in the art.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. The terminology used in thedescription of the invention herein is for the purpose of describingparticular embodiments only and is not intended to be limiting of theinvention.

Unless the context indicates otherwise, it is specifically intended thatthe various features and embodiments of the invention described hereincan be used in any combination.

Moreover, the present invention also contemplates that in someembodiments of the invention, any feature or combination of features setforth herein can be excluded or omitted. To illustrate, if thespecification states that a composition comprises components A, B and C,it is specifically intended that any of A, B or C, or a combinationthereof, can be omitted and disclaimed singularly or in any combination.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. The terminology used in thedescription of the invention herein is for the purpose of describingparticular embodiments only and is not intended to be limiting of theinvention.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Definitions.

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

As used in the description of the invention and the appended claims, thesingular forms “a,” “an” and “the” are intended to include the pluralforms as well, unless the context clearly indicates otherwise.

As used herein, “and/or” refers to and encompasses any and all possiblecombinations of one or more of the associated listed items, as well asthe lack of combinations when interpreted in the alternative (“or”).

The term “about,” as used herein when referring to a measurable valuesuch as a dosage or time period and the like, is meant to encompassvariations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of thespecified amount.

The term “comprise,” “comprises” and “comprising” as used herein,specify the presence of the stated features, integers, steps,operations, elements, and/or components, but do not preclude thepresence or addition of one or more other features, integers, steps,operations, elements, components, and/or groups thereof.

As used herein, the transitional phrase “consisting essentially of”means that the scope of a claim is to be interpreted to encompass thespecified materials or steps recited in the claim “and those that do notmaterially affect the basic and novel characteristic(s)” of the claimedinvention. See, In re Herz, 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463(CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus,the term “consisting essentially of” when used in a claim or thedescription of this invention is not intended to be interpreted to beequivalent to “comprising.”

“Allele”. An allele is any of one or more alternative forms of a gene,all of which relate to a trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

“Backcrossing”. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

“Big Vein virus”. Big vein is a disease of lettuce caused by LettuceMirafiori Big Vein Virus which is transmitted by the fungus Olpidiumvirulentus, with vein clearing and leaf shrinkage resulting in plants ofpoor quality and reduced marketable value.

“Bolting”. The premature development of a flowering stalk, andsubsequent seed, before a plant produces a food crop. Bolting istypically caused by late planting when temperatures are low enough tocause vernalization of the plants.

“Bremia lactucae”. An Oomycete that causes downy mildew in lettuce incooler growing regions.

“Core length”. Length of the internal lettuce stem measured from thebase of the cut and trimmed head to the tip of the stem.

“Corky root”. A disease caused by the bacterium Sphingomonassuberifaciens, which causes the entire taproot to become brown, severelycracked, and non-functional.

“Cotyledon”. One of the first leaves of the embryo of a seed plant;typically one or more in monocotyledons, two in dicotyledons, and two ormore in gymnosperms.

“Double haploid line”. A stable inbred line achieved by doubling thechromosomes of a haploid line, e.g., from anther culture. For example,some pollen grains (haploid) cultivated under specific conditionsdevelop plantlets containing 1 n chromosomes. The chromosomes in theseplantlets are then induced to “double” (e.g., using chemical means)resulting in cells containing 2n chromosomes. The progeny of theseplantlets are termed “double haploid” and are essentially notsegregating any more (e.g., are stable). The term “double haploid” isused interchangeably herein with “dihaploid.”

“Essentially all of the physiological and morphologicalcharacteristics”. A plant having “essentially all of the physiologicaland morphological characteristics” (and similar phrases) means a planthaving all of the physiological and morphological characteristics oflettuce cultivar Chalone, except for the characteristic(s) derived froma converted locus/loci (e.g., a single converted locus), for example,introduced via backcrossing to variety Chalone, a modified gene(s)resulting from genome editing techniques, an introduced transgene (i.e.,introduced via genetic transformation techniques) or mutation, when bothplants are grown under the same environmental conditions. Inembodiments, a plant having “essentially all of the physiological andmorphological characteristics” means a plant having all of thecharacteristics of the reference plant with the exception of five orfewer traits, 4 or fewer traits, 3 or fewer traits, 2 or fewer traits,or one trait. In embodiments, a plant having “essentially all of thephysiological and morphological characteristics” (and similar phrases)optionally has heavy head weight and large head circumference.

“First water date”. The date the seed first receives adequate moistureto germinate. This can and often does equal the planting date.

“Gene”. As used herein, “gene” refers to a segment of nucleic acidcomprising an open reading frame. A gene can be introduced into a genomeof a species, whether from a different species or from the same species,using transformation or various breeding methods.

“Head diameter”. Diameter of the cut and trimmed head, slicedvertically, and measured at the widest point perpendicular to the stem.

“Head height”. Height of the cut and trimmed head, sliced vertically,and measured from the base of the cut stem to the cap leaf.

“Head weight”. Weight of saleable lettuce head, cut and trimmed tomarket specifications.

“Inbred line”: As used herein, the phrase “inbred line” refers to agenetically homozygous or nearly homozygous population. An inbred line,for example, can be derived through several cycles of sib crossingand/or selfing and/or via double haploid production. In someembodiments, inbred lines breed true for one or more traits of interest.An “inbred plant” or “inbred progeny” is an individual sampled from aninbred line.

“Lettuce Mosaic virus”. A disease that can cause a stunted, deformed, ormottled pattern in young lettuce and yellow, twisted, and deformedleaves in older lettuce.

“Maturity date”. Maturity refers to the stage when the plants are offull size and/or optimum weight and/or in marketable form to be ofcommercial or economic value.

“Nasonovia ribisnigri”. A lettuce aphid that colonizes the innermostleaves of the lettuce plant, contaminating areas that cannot be treatedeasily with insecticides.

“Plant.” As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,ovules, seeds, fruit, stems, and the like.

“Plant material”. The terms “plant material” and “material obtainablefrom a plant” are used interchangeably herein and refer to any plantmaterial obtainable from a plant including without limitation, leaves,stems, roots, flowers or flower parts, fruits, pollen, ovules, zygotes,seeds, cuttings, cell or tissue cultures, or any other part or productof the plant.

“Plant part”. As used herein, a “plant part” includes any part, organ,tissue or cell of a plant including without limitation an embryo,meristem, leaf, pollen, cotyledon, hypocotyl, root, root tip, anther,flower, flower bud, pistil, ovule, seed, shoot, stem, stalk, petiole,pith, capsule, a scion, a rootstock and/or a fruit including callus andprotoplasts derived from any of the foregoing.

“Quantitative Trait Loci”. Quantitative Trait Loci (QTL) refers togenetic loci that control to some degree, numerically representabletraits that are usually continuously distributed.

“Ratio of head height/diameter”. Head height divided by the headdiameter is an indication of the head shape; <1 is flattened, 1=round,and >1 is pointed.

“Regeneration”. Regeneration refers to the development of a plant fromtissue culture.

“Resistance”. As used herein the terms “resistance” and “tolerance” (andgrammatical variations thereof) are used interchangeably to describeplants that show reduced or essentially no symptoms to a specific biotic(e.g., a pest, pathogen or disease) or abiotic (e.g., exogenous orenvironmental, including herbicides) factor or stressor. In someembodiments, “resistant” or “tolerant” plants show some symptoms but arestill able to produce marketable product with an acceptable yield, e.g.,the yield may still be reduced and/or the plants may be stunted ascompared with the yield or growth in the absence of the biotic and/orabiotic factor or stressor. Those skilled in the art will appreciatethat the degree of resistance or tolerance may be assessed with respectto a plurality or even an entire field of plants. A lettuce plant may beconsidered “resistant” or “tolerant” if resistance/tolerance is observedover a plurality of plants (e.g., an average), even if particularindividual plants may be susceptible to the biotic or abiotic factor orstressor.

“RHS”. RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd., RHS Garden;Wisley, Woking; Surrey GU236QB, UK.

“Single locus converted”. A single locus converted or conversion plantrefers to a plant that is developed by plant breeding techniques (e.g.,backcrossing), genome editing techniques, genetic transformationtechniques and/or mutation techniques wherein essentially all of thedesired morphological and physiological characteristics of a line arerecovered in addition to the single locus introduced into the line viathe plant breeding, genome editing, genetic transformation, or mutationtechniques.

“Substantially equivalent characteristic”. A characteristic that, whencompared, does not show a statistically significant difference (e.g.,p=0.05) from the mean.

“Tip burn”. Means a browning of the edges or tips of lettuce leaves thatis a physiological response to a lack of calcium.

“Tomato Bushy Stunt”. Also called “lettuce necrotic stunt”. A diseasethat causes stunting of growth and leaf mottling.

“Transgene”. A nucleic acid of interest that can be introduced into thegenome of a plant by genetic engineering techniques (e.g.,transformation) or breeding. The transgene can be from the same or adifferent species. If from the same species, the transgene can be anadditional copy of a native coding sequence or can present the nativesequence in a form or context (e.g., different genomic location and/orin operable association with exogenous regulatory elements such as apromoter) than is found in the native state. The transgene can comprisean open reading frame encoding a polypeptide or can encode a functionalnon-translated RNA (e.g., RNAi).

Botanical Description of the Lettuce Cultivar Chalone (LS17804).

Characteristics. Lettuce cultivar Chalone (LICU18-0048) is an Iceberglettuce variety suitable for late spring to fall production in coastalCalifornia. Lettuce variety Chalone resulted from a cross of Iceberglettuce varieties and several generations of individual plant selectionschosen for head shape, bottom quality, head size, and head weight.

Lettuce variety Chalone (LICU18-0048) has shown uniformity and stabilityfor the traits, within the limits of environmental influence. It hasbeen self-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The variety has been increasedwith continued observation for uniformity. No variant traits have beenobserved or are expected in variety Chalone (LICU18-0048).

TABLE 1 Variety Description Information Plant Type: Iceberg Seed Seedcolor: Black Light dormancy: Light not required Heat dormancy: Nottested Cotyledon to Fourth Leaf Stage Shape of cotyledons: SpatulateShape of 4^(th) Leaf: Obovate Apical Margin: Crenate Basel Margin:Finely Dentate Undulation: Medium Green color: Light green Anthocyanindistribution: Absent Rolling: Present Cupping: Slight Reflexing: NoneMature Leaves Margin incision depth: Absent Margin indentation:Shallowly Dentate Margin undulation of the apical margin: Moderate Greencolor of outer leaves: Light Green Anthocyanin distribution: AbsentGlossiness: Moderate Leaf blistering: Absent/Slight Trichomes: AbsentLeaf thickness: Thin Plant at Market Stage Head shape: SlightlyFlattened Head size class: Medium Head weight (g): 760.08 Head firmness:Moderate Butt shape: Rounded Butt Midrib: Moderately Raised CoreDiameter at base of head (mm): 31.32 Core height from base of head toapex (mm): 55.25 Maturity (days) Spring: 65 to 85 days Summer: 65 to 75days Fall: 65 to 75 days Winter: Not Adapted Adaptation Primary U.S.Regions of Adaptation (tested and proven adapted) Southwest (California,Arizona desert): Not Adapted West Coast: Yes Southeast: Not testedNortheast: Not tested Spring area: West Coast Summer area: West CoastFall area: West Coast Winter area: Not Adapted Greenhouse: Not testedSoil type: Both of mineral and organic Plant habit at bolting Boltingclass: Medium Bolter leaves: Curved Leaf margin: Entire Leaf color:Medium green Terminal inflorescence: Present Lateral shoots: PresentBasel side shoots: Absent Disease and Stress Reactions Virus TomatoBushy Stunt virus (TBSV): Highly resistant Big vein: Not tested LettuceMosaic: Not tested Cucumber Mosaic virus: Not tested Broad Bean Wilt:Not tested Turnip Mosaic virus: Not tested Best Western Yellows: Nottested Lettuce Infectious Yellows: Not tested Lettuce Mosaic Virusstrain Ls-1: Susceptible Fungal/Bacterial Corky Root Rot (Pythium RootRot): Not tested Bremia lactucae (Downy Mildew): Highly Resistant: Bl:16, 21, 23, 32EU, Bl: 5-6US Powdery Mildew: Not tested Sclerotinia Rot:Not tested Bacterial Soft Rot (Pseudomonas spp. & others): Not testedBotrytis (Gray Mold): Not tested Insects Cabbage Loopers: Not testedNasonovia ribisnigri biotype Nr:0: Susceptible Root Aphids: Not testedGreen Peach Aphid: Not tested Physiological/Stress Tip burn: Not testedHeat: Intermediate Drought: Not tested Cold: Not Adapted Salt: Nottested Brown Rib: Not tested Post-Harvest Pink Rib: Not tested RussettSpotting: Not tested Rusty Brown Discoloration: Not tested Internal RibNecrosis (Blackheart, Gray Rib, Gray Streak): Not tested Brown Stain:Not tested

TABLE 2 Distinguishing characteristics of cultivar ChaloneCharacteristic Plant Stage Large frame leaf Harvest Mature length towidth index Stage Large head circumference Medium core length Narrowcore width Narrow frame leaf width Heavy head weight Large core lengthto width index Short fourth leaf Young Plant Stage length (20 days)Short fourth leaf width Medium Cotyledon width

TABLE 3 Length (mm) of fourth leaf at 20 Days Chalone Telluride Reliant30 20 34 27 21 20 30 19 23 33 23 21 25 20 34 14 17 20 13 30 35 20 27 2220 19 40 20 23 35 14 38 38 15 28 25 13 15 32 20 15 37 28 14 24 14 17 2515 16 28 14 27 27 22 20 24 19 19 29 Response Length (mm) of fourth leafat 20 ANOVA Variable: Days Source of Variation df SS MS F value Pr (>F)Variety 2 823.30 411.7 10.275 <0.001 Residuals 57 2283.55 40.1 VarietyMean Duncan Grouping Reliant 28.65 a Telluride 21.40 b Chalone 20.30 bANOVA shows a significant difference (p < 0.05) in the length (mm) offourth leaf at 20 days. The average length (mm) of fourth leaf forReliant, Telluride, and Chalone is 28.65, 21.40, and 20.30,respectively. However, based on the Duncan Grouping, there are nodifference between means for Telluride and Chalone.

TABLE 4 Width (mm) of fourth Leaf at 20 Days Chalone Telluride Reliant18 15 19 17 16 11 20 10 18 18 16 15 18 17 20 10 12 15 9 15 18 19 20 13 910 20 12 14 22 8 18 22 9 17 14 8 10 19 11 12 19 16 11 14 10 10 17 11 818 11 15 17 12 16 16 14 12 17 Response Width (mm) of fourth Leaf ANOVAVariable: at 20 Days Source of Variation df SS MS F value Pr (>F)Variety 2 202.533 101.27 8.471 0.001 Residuals 57 681.400 11.95 VarietyMean Duncan Grouping Reliant 17.2 a Telluride 13.7 b Chalone 13.0 bANOVA shows a significant difference (p < 0.05) in fourth leaf width(mm) at 20 days, and the average width (mm) for the fourth leaf forReliant, Telluride, and Chalone is 17.2, 13.7, and 13.0, respectively.However, based on the Duncan Grouping, there are no differences betweenmeans for Telluride and Chalone.

TABLE 5 Length to Width Index of fourth Leaf at 20 Days ChaloneTelluride Reliant 1.67 1.33 1.79 1.59 1.31 1.82 1.50 1.90 1.28 1.83 1.441.40 1.39 1.18 1.70 1.40 1.42 1.33 1.44 2.00 1.94 1.05 1.35 1.69 2.221.90 2.00 1.67 1.64 1.59 1.75 2.11 1.73 1.67 1.65 1.79 1.63 1.50 1.681.82 1.25 1.95 1.75 1.27 1.71 1.40 1.70 1.47 1.36 2.00 1.56 1.27 1.801.59 1.83 1.25 1.50 1.36 1.58 1.71 Response Length to Width Index ofANOVA Variable: fourth Leaf at 20 Days Source of Variation df SS MS Fvalue Pr (>F) Variety 2 0.090 0.045 0.699 0.501 Residuals 57 3.654 0.063Variety Mean Duncan Grouping Reliant 1.66 a Chalone 1.58 a Telluride1.58 a ANOVA shows no significant difference (p < 0.05) in length towidth index of fourth leaf at 20 Days.

TABLE 6 Length (mm) of Cotyledon at 20 Days Chalone Telluride Reliant 1717 20 17 20 19 16 19 20 15 18 17 18 17 17 20 18 18 18 17 17 18 15 16 1717 20 16 16 21 19 15 22 21 20 20 18 18 17 16 17 21 18 20 17 15 18 17 1820 17 20 17 15 20 17 16 19 16 18 Response Length (mm) of Cotyledon ANOVAVariable: at 20 Days Source of Variation df SS MS F value Pr (>F)Variety 2 4.422 2.217 0.719 0.492 Residuals 57 175.750 3.083 VarietyMean Duncan Grouping Reliant 18.25 a Chalone 17.80 a Telluride 17.60 aANOVA shows no significant difference (p < 0.05) in cotyledon length(mm) at 20 days among Reliant, Telluride, and Chalone.

TABLE 7 Width (mm) of Cotyledon at 20 Days Chalone Telluride Reliant 9 99 9 10 10 9 10 9 8 8 9 9 9 9 10 9 10 9 9 9 9 9 9 8 8 11 7 7 10 10 8 1210 9 10 9 9 10 9 9 10 9 9 9 8 9 10 9 10 9 10 9 10 10 9 9 9 8 10 ResponseWidth (mm) of Cotyledon ANOVA Variable: at 20 Days Source of Variationdf SS MS F value Pr (>F) Variety 2 8.233 4.117 6.753 0.002 Residuals 5734.750 0.610 Variety Mean Duncan Grouping Reliant 9.70 a Chalone 9.00 bTelluride 8.85 b ANOVA shows a significant difference (p < 0.01) incotyledon width (mm) at 20 days and the average width (mm) for thecotyledon for Reliant, Chalone, and Telluride is 9.70, 9.00, and 8.85,respectively. However, based on the Duncan Grouping, there are nodifferences between means for Telluride and Chalone.

TABLE 8 Length to Width Index of Cotyledon Leaf at 20 Days ChaloneTelluride Reliant 1.89 1.89 2.22 1.89 2.00 1.90 1.78 1.90 2.22 1.88 2.251.89 2.00 1.89 1.89 2.00 2.00 1.80 2.00 1.89 1.89 2.00 1.67 1.78 2.132.13 1.82 2.29 2.29 2.10 1.90 1.88 1.83 2.10 2.22 2.00 2.00 2.00 1.701.78 1.89 2.10 2.00 2.22 1.89 1.88 2.00 1.70 2.00 2.00 1.89 2.00 1.891.50 2.00 1.89 1.78 2.11 2.00 1.80 Response Length to Width Index ofANOVA Variable: Cotyledon Leaf at 20 Days Source of Variation df SS MS Fvalue Pr (>F) Variety 2 0.144 0.072 3.079 0.054 Residuals 57 1.333 0.023Variety Mean Duncan Grouping Telluride 1.99 a Chalone 1.98 ab Reliant1.88 b ANOVA shows a significant difference (p < 0.10) in Length toWidth index of Cotyledon Leaf at 20 Days and the average of the indexfor Telluride, Chalone, and Reliant is 1.98, 1.98, and 1.88,respectively. However, based on the Duncan Grouping, there are nodifferences between means for Telluride and Chalone and the means forChalone and Reliant.

TABLE 9 Head Weight (g) at Harvest Maturity Location 1 Location 2Chalone Reliant Telluride Chalone Reliant Telluride 633 790 964 617 519742 748 1009 840 664 427 627 790 633 720 611 753 675 1126 674 897 732640 626 677 918 635 1008 640 867 841 743 661 903 676 699 688 896 760 945365 758 466 799 694 706 727 748 521 648 686 1128 505 575 518 915 7011081 442 936 727 612 606 792 579 672 696 796 538 731 515 896 588 774 413665 632 872 654 727 653 775 525 501 881 800 353 820 473 658 491 533 726983 796 713 755 516 517 730 530 653 631 537 409 942 421 738 771 596 391885 591 780 821 885 559 662 405 726 Response Head Weight (g) ANOVAVariable: at Harvest Maturity Source of Variation df SS MS F value Pr(>F) Variety 2 263114.5 131540 6.376 0.002 Location 1 1732.8 1724 0.0840.773 Variety:Location 2 543996.4 272097 13.182 <0.001 Residuals 1142352322.8 20637 Variety Mean Duncan Grouping Chalone 760.08 a Telluride679.62 b Reliant 649.05 b ANOVA shows a significant difference (p <0.01) for head weight (g) at harvest mature stage for variety and in theinteraction between variety and location. The average head weight (g) ofChalone, Telluride, and Reliant are 760.08, 679.62, and 649.05,respectively. Based on the Duncan Grouping, there are no differencesbetween means for Telluride and Reliant.

TABLE 10 Head Circumference (cm) at Harvest Maturity Location 1 Location2 Chalone Reliant Telluride Chalone Reliant Telluride 52 52 56 58 55 6155 55 51 54 53 53 52 52 52 54 57 52 57 50 55 56 54 55 53 56 49 57 53 5755 53 47 62 53 58 52 55 48 61 46 54 45 55 47 53 53 56 46 52 48 58 54 5349 53 48 59 46 57 52 48 48 55 54 53 51 51 47 55 53 55 51 52 45 55 52 5851 51 48 56 53 57 57 49 42 54 47 54 48 47 52 52 54 54 57 45 44 54 56 5652 47 45 56 51 55 57 48 42 58 54 60 56 57 46 51 56 54 Response HeadCircumference (cm) ANOVA Variable: at Harvest Maturity Source ofVariation df SS MS F value Pr (>F) Variety 2 133.267 66.6 6.524 0.002Location 1 512.533 512.5 50.179 <0.001 Variety:Location 2 204.467 102.210.009 <0.001 Residuals 114 1164.400 10.2 Variety Mean Duncan GroupingChalone 54.15 a Reliant 52.05 b Telluride 51.80 b ANOVA shows asignificant difference (p < 0.001) in head circumference (cm) at harvestmature stage for variety, location, and in the interaction betweenvariety and location. The average head circumference (cm) of Chalone,Reliant, and Telluride are 54.15, 52.05, and 51.08, respectively. Basedon the Duncan Grouping, there are no differences between means forTelluride and Reliant.

TABLE 11 Core Length (mm) at Harvest Maturity Location 1 Location 2Chalone Reliant Telluride Chalone Reliant Telluride 55 57 71 58 29 60 7570 62 57 32 58 50 44 57 56 38 60 68 60 59 64 23 52 48 44 70 50 31 62 6262 71 63 23 50 55 61 68 62 27 60 48 56 52 60 20 51 45 51 60 52 29 52 5060 76 65 19 52 60 52 94 62 32 60 50 60 62 70 40 57 43 50 62 48 28 54 5859 62 51 40 61 58 73 48 52 32 50 45 50 67 68 32 65 56 45 52 40 40 60 4652 59 47 45 52 49 49 56 49 23 46 65 55 62 50 35 55 Response Core Length(mm) ANOVA Variable: at Harvest Maturity Source of Variation df SS MS Fvalue Pr (>F) Variety 2 5816.117 2908.1 45.982 <0.001 Location 13070.408 3070.4 48.549 <0.001 Variety:Location 2 3602.517 1801.3 28.481<0.001 Residuals 114 7209.750 63.2 Variety Mean Duncan GroupingTelluride 59.67 a Chalone 55.25 b Reliant 43.20 c ANOVA shows asignificant difference (p < 0.05) in core length (mm) at harvest maturestage for variety, location, and in the interaction between variety andlocation. The average core length (mm) of Telluride, Chalone, andReliant are 59.67, 55.25, and 43.20, respectively.

TABLE 12 Core Width (mm) at Harvest Maturity Location 1 Location 2Chalone Reliant Telluride Chalone Reliant Telluride 29 29 36 33 35 38 3034 30 32 37 32 30 28 33 32 38 40 33 29 34 39 28 35 29 29 32 31 39 39 3030 35 32 34 32 28 32 35 34 35 36 25 29 34 25 33 36 24 29 29 33 40 36 2930 32 35 30 32 30 29 30 35 37 34 30 32 30 35 38 38 29 30 28 36 34 33 3028 33 39 38 39 30 31 26 34 36 38 27 24 30 36 36 38 31 25 29 36 35 40 3026 29 27 38 35 30 24 29 33 33 37 30 30 30 32 36 40 Response Core Width(mm) ANOVA Variable: at Harvest Maturity Source of Variation df SS MS Fvalue Pr (>F) Variety 2 126.017 63.0 8.266 <0.001 Location 1 858.675858.7 112.652 <0.001 Variety:Location 2 27.950 14.0 1.833 0.165Residuals 114 868.950 7.6 Variety Mean Duncan Grouping Telluride 33.80 aReliant 32.20 b Chalone 31.32 b ANOVA shows a significant difference (p< 0.001) in core width (mm) at harvest mature stage for variety andlocation. The average core width (mm) of Telluride, Reliant, and Chaloneare 33.80, 32.20, and 31.32, respectively. Based on the Duncan Grouping,there are no differences between means for Reliant and Chalone.

TABLE 13 Frame Leaf Length (cm) at Harvest Maturity Location 1 Location2 Chalone Reliant Telluride Chalone Reliant Telluride 27 29 23 31 31 2727 27 26 26 28 26 26 25 29 26 29 27 25 26 31 25 32 30 27 28 26 25 32 2728 24 31 29 30 27 27 30 27 25 31 26 27 27 21 29 32 26 28 22 30 30 28 2723 25 27 28 29 29 23 26 26 26 30 28 27 24 27 24 26 25 27 29 26 31 30 2723 28 30 25 26 32 26 27 24 29 25 23 26 26 26 28 29 26 26 25 27 29 30 3027 23 25 30 27 27 27 23 27 30 27 25 27 24 26 29 27 25 Response FrameLeaf Length (cm) ANOVA Variable: at Harvest Maturity Source of Variationdf SS MS F value Pr (>F) Variety 2 6.867 3.43 0.742 0.478 Location 178.408 78.41 16.947 <0.001 Variety:Location 2 39.267 19.63 4.243 0.017Residuals 114 527.450 4.63 Variety Mean Duncan Grouping Reliant 27.43 aChalone 26.98 a Telluride 26.88 a ANOVA shows no significant difference(p < 0.05) in frame leaf length (cm) for varieties, but there wassignificant difference (p < 0.001) between locations and in theinteraction between variety and location.

TABLE 14 Frame Leaf Width (cm) at Harvest Maturity Location 1 Location 2Chalone Reliant Telluride Chalone Reliant Telluride 25 29 30 28 36 29 2727 30 29 36 28 26 29 30 31 35 29 25 30 27 29 35 34 29 23 23 32 38 32 2526 31 28 42 31 25 27 31 24 39 28 25 25 26 33 36 29 25 27 33 34 35 26 2525 30 28 35 31 23 28 30 31 35 29 30 27 31 25 31 29 25 29 28 34 37 28 2227 28 29 35 31 25 30 25 30 36 28 26 31 25 28 35 28 27 25 30 34 36 29 2626 26 29 35 28 25 28 33 34 34 28 26 25 29 29 38 26 Response Frame LeafWidth (cm) ANOVA Variable: at Harvest Maturity Source of Variation df SSMS F value Pr (>F) Variety 2 303.800 151.9 28.667 <0.001 Location 1594.075 594.1 112.117 <0.001 Variety:Location 2 361.400 180.7 34.103<0.001 Residuals 114 604.050 5.3 Variety Mean Duncan Grouping Reliant31.57 a Telluride 28.93 b Chalone 27.77 c ANOVA shows a significantdifference (p < 0.001) in frame leaf width (cm) at harvest mature stagefor variety, location, and in the interaction between variety andlocation. The average frame leaf width (cm) of Chalone, Reliant, andTelluride are 31.57, 28.93, and 27.77, respectively.

TABLE 15 Core length to width index at Harvest Maturity Location 1Location 2 Chalone Reliant Telluride Chalone Reliant Telluride 1.90 1.971.97 1.76 0.83 1.58 2.50 2.06 2.07 1.78 0.86 1.81 1.67 1.57 1.73 1.751.00 1.50 2.06 2.07 1.74 1.64 0.82 1.49 1.66 1.52 2.19 1.61 0.79 1.592.07 2.07 2.03 1.97 0.68 1.56 1.96 1.91 1.94 1.82 0.77 1.67 1.92 1.931.53 2.40 0.61 1.42 1.88 1.76 2.07 1.58 0.73 1.44 1.72 2.00 2.38 1.860.63 1.63 2.00 1.79 3.13 1.77 0.86 1.76 1.67 1.88 2.07 2.00 1.05 1.501.48 1.67 2.21 1.33 0.82 1.64 1.93 2.11 1.88 1.31 1.05 1.56 1.93 2.351.85 1.53 0.89 1.32 1.67 2.08 2.23 1.89 0.89 1.71 1.81 1.80 1.79 1.111.14 1.50 1.53 2.00 2.03 1.74 1.18 1.49 1.63 2.04 1.93 1.48 0.70 1.242.17 1.83 2.07 1.56 0.97 1.38 Response Core length to width index ANOVAVariable: at Harvest Maturity Source of Variation df SS MS F value Pr(>F) Variety 2 4.091 2.046 37.339 <0.001 Location 1 9.896 9.896 180.621<0.001 Variety:Location 2 4.069 2.035 37.135 <0.001 Residuals 114 6.2460.055 Variety Mean Duncan Grouping Telluride 1.79 a Chalone 1.78 aReliant 1.39 b ANOVA shows a significant difference (p < 0.001) in corelength to width index at harvest mature stage for variety, location, andin the interaction between variety and location. The average core lengthto width index of Telluride, Chalone, and Reliant are 1.79, 1.78, and1.39, respectively. Based on the Duncan Grouping, there are nodifferences between means for Telluride and Chalone.

TABLE 16 Frame Leaf Length to Width Index at Harvest Maturity Location 1Location 2 Chalone Reliant Telluride Chalone Reliant Telluride 1.08 1.000.77 1.11 0.86 0.93 1.00 1.00 0.87 0.90 0.78 0.93 1.00 0.86 0.97 0.840.83 0.93 1.00 0.87 1.15 0.86 0.91 0.88 0.93 1.22 1.13 0.78 0.84 0.841.12 0.92 1.00 1.04 0.71 0.87 1.08 1.11 0.87 1.04 0.79 0.93 1.08 1.080.81 0.88 0.89 0.90 1.12 0.81 0.91 0.88 0.80 1.04 0.92 1.00 0.90 1.000.83 0.94 1.00 0.93 0.87 0.84 0.86 0.97 0.90 0.89 0.87 0.96 0.84 0.861.08 1.00 0.93 0.91 0.81 0.96 1.05 1.04 1.07 0.86 0.74 1.03 1.04 0.900.96 0.97 0.69 0.82 1.00 0.84 1.04 1.00 0.83 0.93 0.96 1.00 0.90 0.850.83 1.03 1.04 0.88 0.96 1.03 0.77 0.96 1.08 0.82 0.82 0.88 0.79 0.891.04 0.96 0.90 1.00 0.71 0.96 Response Frame Leaf Length to Width IndexANOVA Variable: at Harvest Maturity Source of Variation df SS MS F valuePr (>F) Variety 2 0.191 0.095 14.153 <0.001 Location 1 0.209 0.20930.952 <0.001 Variety:Location 2 0.108 0.054 8.020 <0.001 Residuals 1140.770 0.006 Variety Mean Duncan Grouping Chalone 0.98 a Telluride 0.93 bReliant 0.88 c ANOVA shows a significant difference (p < 0.001) in frameleaf length to width index at harvest mature stage for variety,location, and in the interaction between variety and location. Theaverage frame leaf length to width index of Chalone, Telluride, andReliant are 0.98, 0.93, and 0.88, respectively.

Tissue Culture.

Further reproduction of lettuce plants variety can occur by tissueculture and regeneration. Tissue culture of various tissues of lettuceand regeneration of plants therefrom is well known and widely published.For example, reference may be had to Teng, et al., HortScience, 27:9,1030-1032 (1992); Teng, et al., HortScience, 28:6, 669-1671 (1993);Zhang, et al., Journal of Genetics and Breeding, 46:3, 287-290 (1992);Webb, et al., Plant Cell Tissue and Organ Culture, 38:1, 77-79 (1994);Curtis, et al., Journal of Experimental Botany, 45:279, 1441-1449(1994); Nagata, et al., Journal for the American Society forHorticultural Science, 125:6, 669-672 (2000); and Ibrahim, et al., PlantCell Tissue and Organ Culture, 28(2), 139-145 (1992). It is clear fromthe literature that the state of the art is such that these methods ofobtaining plants are routinely used and have a very high rate ofsuccess. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce lettuce plants havingdesired characteristics of lettuce cultivar Chalone. Optionally, lettuceplants can be regenerated from the tissue culture of the inventioncomprising all or essentially all of the physiological and morphologicalcharacteristics of lettuce cultivar Chalone.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers, and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques.

Additional Breeding Methods.

This invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant wherein the first or second parent lettuce plant is a plant oflettuce cultivar Chalone. Further, both first and second parent lettucecan come from lettuce cultivar Chalone. Thus, any of the followingexemplary methods using lettuce cultivar Chalone are part of thisinvention: selfing, backcrosses, hybrid production, crosses topopulations, double haploid production, and the like. All plantsproduced using lettuce cultivar Chalone as at least one parent arewithin the scope of this invention, including those developed fromlettuce plants derived from lettuce cultivar Chalone. Advantageously,lettuce cultivar Chalone can be used in crosses with other, different,lettuce plants to produce the first generation (F₁) lettuce hybrid seedsand plants with desirable characteristics. The lettuce plants of theinvention can also be used for transformation where exogenous transgenesare introduced and expressed by the plants of the invention. Geneticvariants created either through traditional breeding methods or throughtransformation of the cultivars of the invention by any of a number ofprotocols known to those of skill in the art are intended to be withinthe scope of this invention.

The following describes exemplary breeding methods that may be used withlettuce cultivar Chalone in the development of further lettuce plants.One such embodiment is a method for developing lettuce cultivar Chaloneprogeny lettuce plants in a lettuce plant breeding program comprising:obtaining a plant, or a part thereof, of lettuce cultivar Chalone,utilizing said plant or plant part as a source of breeding material, andselecting a lettuce cultivar Chalone progeny plant with molecularmarkers in common with lettuce cultivar Chalone and/or with some, all oressentially all of the morphological and/or physiologicalcharacteristics of lettuce cultivar Chalone (see, e.g., Table 2 andTables 1 and 3-16). In representative embodiments, the progeny plant hasat least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of the morphological andphysiological characteristics of lettuce cultivar Chalone (e.g., asdescribed in Table 2 and Tables 1 and 3-16) or even all of themorphological and physiological characteristics of lettuce cultivarChalone so that said progeny lettuce plant is not significantlydifferent for said traits than lettuce cultivar Chalone, as determinedat the 5% significance level when grown in the same environmentalconditions; optionally, with the presence of one or more desiredadditional traits (e.g., male sterility, disease resistance, pest orinsect resistance, herbicide resistance, and the like). Breeding stepsthat may be used in the breeding program include pedigree breeding,backcrossing, mutation breeding and/or recurrent selection. Inconjunction with these steps, techniques such as RFLP-enhancedselection, genetic marker enhanced selection (for example, SSR markers)and/or and the making of double haploids may be utilized.

Another representative method involves producing a population of lettucecultivar Chalone progeny plants, comprising crossing lettuce cultivarChalone with another lettuce plant, thereby producing a population oflettuce plants that, on average, derives 50% of its alleles (i.e., TAC)from lettuce cultivar Chalone. A plant of this population may beselected and repeatedly selfed or sibbed with a lettuce plant resultingfrom these successive filial generations or backcrossed to lettucecultivar Chalone. Another approach is to make double haploid plants toachieve homozygosity. One embodiment of this invention is a lettuceplant produced by these methods and that has obtained at least 50% ofits alleles from lettuce cultivar Chalone. In embodiments, the methodsof the invention produce a population of lettuce plants that, onaverage, derives at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of itsalleles (i.e., TAC) from lettuce cultivar Chalone, e.g., at least about6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%,85%, 90%, 95%, 96%, 97%, 98% or 99% of the genetic complement of lettucecultivar Chalone. One representative embodiment of this invention is thelettuce plant produced by the methods of the invention and that hasobtained at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles (i.e.,TAC) from lettuce cultivar Chalone, and optionally is the result of abreeding process comprising one or two breeding crosses and one or moreof selfing, sibbing, backcrossing and/or double haploid techniques inany combination and any order. In embodiments, the breeding process doesnot include a breeding cross, and comprises selfing, sibbing,backcrossing and or double haploid technology.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr and Walt, Principles of CultivarDevelopment, pp. 261-286 (1987). In embodiments, the inventionencompasses Chalone progeny plants having a combination of at least 2,3, 4, 5, 6, 7, 8, 9, 10 or more of the characteristics as describedherein for lettuce cultivar Chalone, so that said progeny lettuce plantis not significantly different for said traits than lettuce cultivarChalone, as determined at the 5% significance level when grown in thesame environmental conditions. Using techniques described herein andthose known in the art, molecular markers may be used to identify saidprogeny plant as progeny of lettuce cultivar Chalone. Mean trait valuesmay be used to determine whether trait differences are significant, andoptionally the traits are measured on plants grown under the sameenvironmental conditions.

Progeny of lettuce cultivar Chalone may also be characterized throughtheir filial relationship with lettuce cultivar Chalone, as for example,being within a certain number of breeding crosses of lettuce cultivarChalone. A breeding cross is a cross made to introduce new genetics intothe progeny, and is distinguished from a cross, such as a self or a sibcross or a backcross to Chalone as a recurrent parent, made to selectamong existing genetic alleles. The lower the number of breeding crossesin the pedigree, the closer the relationship between lettuce cultivarChalone and its progeny. For example, progeny produced by the methodsdescribed herein may be within 1, 2, 3, 4, 5 or more breeding crosses oflettuce cultivar Chalone.

In representative embodiments, a lettuce plant derived from lettucecultivar Chalone comprises cells comprising at least one set ofchromosomes derived from lettuce cultivar Chalone. In embodiments, thelettuce plant or population of lettuce plants derived from lettucecultivar Chalone comprises, on average, at least 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of its alleles (i.e., TAC) from lettuce cultivar Chalone,e.g., at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the geneticcomplement of lettuce cultivar Chalone, and optionally is the result ofa breeding process comprising one or two breeding crosses and one ormore of selfing, sibbing, backcrossing and/or double haploid techniquesin any combination and any order. In embodiments, the breeding processdoes not include a breeding cross, and comprises selfing, sibbing,backcrossing and or double haploid technology. In embodiments, thelettuce plant derived from lettuce cultivar Chalone is one, two, three,four, five or more breeding crosses removed from lettuce cultivarChalone.

In representative embodiments, a plant derived from lettuce cultivarChalone is a double haploid plant, a hybrid plant or an inbred plant.

In embodiments, a hybrid or derived plant from lettuce cultivar Chalonecomprises a desired added trait. In representative embodiments, alettuce plant derived from lettuce cultivar Chalone comprises all of themorphological and physiological characteristics of lettuce cultivarChalone (e.g., as described in Table 2 and Tables 1 and 3-16). Inembodiments, the lettuce plant derived from lettuce cultivar Chalonecomprises essentially all of the morphological and physiologicalcharacteristics of lettuce cultivar Chalone (e.g., as described in Table2 and Tables 1 and 3-16) in any combination, with the addition of adesired added trait.

Those skilled in the art will appreciate that any of the traitsdescribed above with respect to plant transformation methods can beintroduced into a plant of the invention (e.g., lettuce cultivar Chaloneand hybrid lettuce plants and other lettuce plants derived therefrom)using breeding techniques.

Genetic Transformation.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign nucleic acids including additional or modified versions ofnative (endogenous) nucleic acids (optionally driven by a non-nativepromoter) in order to alter the traits of a plant in a specific manner.Any nucleic acid sequences, whether from a different species, the samespecies or an artificial sequence, which are introduced into the genomeusing transformation or various breeding methods, are referred to hereincollectively as “transgenes.” Over the last fifteen to twenty years,several methods for producing transgenic plants have been developed, andin particular embodiments the present invention also relates totransformed versions of the plants disclosed herein.

Genetic engineering techniques can be used (alone or in combination withbreeding methods) to introduce one or more desired added traits intoplant, for example, lettuce cultivar Chalone or progeny or lettuceplants derived thereof. Once a transgene has been introduction into aplant by genetic transformation, it can be transferred to other plantsvia conventional breeding.

Plant transformation generally involves the construction of anexpression vector that will function in plant cells. Optionally, such avector comprises one or more nucleic acids comprising a coding sequencefor a polypeptide or an untranslated functional RNA under control of, oroperatively linked to, a regulatory element (for example, a promoter).In representative embodiments, the vector(s) may be in the form of aplasmid, and can be used alone or in combination with other plasmids, toprovide transformed lettuce plants using transformation methods asdescribed herein to incorporate transgenes into the genetic material ofthe lettuce plant.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct nucleic acid transfermethod, such as microprojectile-mediated delivery (e.g., with abiolistic device), DNA injection, Agrobacterium-mediated transformation,electroporation, and the like. Transformed plants obtained from theplants (and parts and tissue culture thereof) of the invention areintended to be within the scope of this invention.

Expression Vectors for Plant Transformation—Selectable markers.

Expression vectors typically include at least one nucleic acidcomprising or encoding a selectable marker, operably linked to aregulatory element (for example, a promoter) that allows transformedcells containing the marker to be either recovered by negativeselection, e.g., inhibiting growth of cells that do not contain theselectable marker, or by positive selection, e.g., screening for theproduct encoded by the selectable marker. Many commonly used selectablemarkers for plant transformation are well known in the transformationart, and include, for example, nucleic acids that code for enzymes thatmetabolically detoxify a selective chemical agent which may be anantibiotic or an herbicide, or nucleic acids that encode an alteredtarget which is insensitive to the inhibitor. Positive selection methodsare also known in the art.

Commonly used selectable markers in plants include, but are not limitedto: neomycin phosphotransferase II (nptII) conferring resistance tokanamycin, hygromycin phosphotransferase conferring resistance to theantibiotic hygromycin, bacterial selectable markers that conferresistance to antibiotics (e.g., gentamycin acetyl transferase,streptomycin phosphotransferase, and aminoglycoside-3′-adenyltransferase, selectable markers conferring resistance to herbicides(e.g., glyphosate, glufosinate, or bromoxynil). Selection of transformedplant cells can also be based on screening presumptively transformedplant cells rather than direct genetic selection of transformed cellsfor resistance to a toxic substance such as an antibiotic; such markersinclude without limitation alpha-glucuronidase (GUS),alpha-galactosidase, luciferase, and Green Fluorescent Protein (GFP) andmutant GFPs.

Expression Vectors for Plant Transformation—Promoters.

Transgenes included in expression vectors are generally driven by anucleotide sequence comprising a regulatory element (for example, apromoter). Numerous types of promoters are well known in thetransformation arts, as are other regulatory elements that can be usedalone or in combination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells.

Examples of promoters under developmental control include promoters thatpreferentially initiate transcription in certain tissues, such asleaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma.Such promoters are referred to as “tissue-preferred.” Promoters thatinitiate transcription only in certain tissue are referred to as“tissue-specific.” A “cell type” specific promoter preferentially drivesexpression in certain cell types in one or more organs, for example,vascular cells in roots or leaves. An “inducible” promoter is a promoterthat is under environmental control. Examples of environmentalconditions that may affect transcription by inducible promoters includeanaerobic conditions or the presence of light. Tissue-specific,tissue-preferred, cell type specific, and inducible promoters constitutethe class of “non-constitutive” promoters. A “constitutive” promoter isa promoter that is active under most environmental conditions.

Many suitable promoters are known in the art and can be selected andused to achieve the desired outcome.

Signal Sequences for Targeting Proteins to Subcellular Compartments.

Transport of polypeptides produced by transgenes to a subcellularcompartment such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isgenerally accomplished by means of operably linking a nucleotidesequence encoding a signal sequence to the 5′ and/or 3′ region of anucleic acid encoding the polypeptide of interest. Signal sequences atthe 5′ and/or 3′ end of the coding sequence target the polypeptide toparticular subcellular compartments.

The presence of a signal sequence can direct a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Close, P. S.,Master's Thesis, Iowa State University (1993); Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., PlantPhysiol., 91:124-129 (1989); Fontes, et al., Plant Cell, 3:483-496(1991); Matsuoka, et al., PNAS, 88:834 (1991); Gould, et al., J. Cell.Biol., 108:1657 (1989); Creissen, et al., Plant J, 2:129 (1991);Kalderon, et al., A short amino acid sequence able to specify nuclearlocation, Cell, 39:499-509 (1984); and Steifel, et al., Expression of amaize cell wall hydroxyproline-rich glycoprotein gene in early leaf androot vascular differentiation, Plant Cell, 2:785-793 (1990).

Foreign Polypeptide Transgenes and Agronomic Transgenes.

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign polypeptide then canbe extracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981). According to a representative embodiment, the transgenic plantprovided for commercial production of foreign protein is a lettuce plantof the invention. In another embodiment, the biomass of interest is seedand/or fruit.

Likewise, by means of the present invention, agronomic transgenes andother desired added traits can be expressed in transformed plants (andtheir progeny, e.g., produced by breeding methods). More particularly,plants can be genetically engineered to express various phenotypes ofagronomic interest or other desired added traits. Exemplary nucleicacids of interest in this regard conferring a desired added trait(s)include, but are not limited to, those transgenes that confer resistanceto confer resistance to plant pests (e.g., nematode or insect) ordisease (e.g., fungal, bacterial or viral), transgenes that conferherbicide tolerance, transgenes that confer male sterility, andtransgenes that confer or contribute to a value-added trait such asincreased nutrient content (e.g., iron, nitrate), increased sweetness(e.g., by introducing a transgene coding for monellin), modified fattyacid metabolism (for example, by introducing into a plant an antisensesequence directed against stearyl-ACP desaturase to increase stearicacid content of the plant), modified carbohydrate composition (e.g., byintroducing into plants a transgene coding for an enzyme that alters thebranching pattern of starch), modified fruit color (e.g., external fruitcolor and/or fruit flesh), or modified flavor profile of the fruit.

In embodiments, the transgene encodes a non-translated RNA (e.g., RNAi)that is expressed to produce targeted inhibition of gene expression,thereby conferring the desired trait on the plant.

In embodiments, the transgene encodes the machinery used for geneediting techniques.

Any transgene, including those exemplified above, can be introduced intothe lettuce plants of the invention through a variety of meansincluding, but not limited to, transformation (e.g., genetic engineeringtechniques), conventional breeding, and introgression methods tointroduce the transgene into other genetic backgrounds.

Methods for Plant Transformation.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993). Commonly usedplant transformation methods include agrobacterium-mediatedtransformation and direct transgene transfer methods (e.g.,microprojectile-mediated transformation, sonication, liposome orspheroplast fusion, and electroporation of protoplasts or whole cells).

Following transformation of plant target tissues, expression ofselectable marker transgenes (e.g., as described above) allows forpreferential selection of transformed cells, tissues and/or plants,using regeneration and selection methods now well known in the art.

The foregoing methods for transformation are typically used to produce atransgenic lettuce line. The transgenic lettuce line can then be crossedwith another (non-transgenic or transgenic) line in order to produce anew transgenic lettuce line. Alternatively, a transgene that has beenengineered into a particular plant using transformation techniques canbe introduced into another plant or line using traditional breeding(e.g., backcrossing) techniques that are well known in the plantbreeding arts. For example, a backcrossing approach can be used to movean engineered transgene from a public, non-elite inbred line into anelite inbred line, or from an inbred line containing a foreign transgenein its genome into an inbred line or lines which do not contain thattransgene. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context.

Locus Conversions.

When the term “lettuce plant” is used in the context of the presentinvention, this term also includes any locus conversions of that plantor variety. The term “locus converted plant” as used herein refers tothose plants that are developed, for example, by backcrossing, genomeediting, genetic transformation and/or mutation, wherein essentially allof the desired morphological and physiological characteristics of avariety are recovered in addition to the one or more loci introducedinto the variety. To illustrate, backcrossing methods can be used withthe present invention to improve or introduce a characteristic into thevariety. The term “backcrossing” as used herein refers to the repeatedcrossing of a hybrid progeny back to the recurrent parent, e.g.,backcrossing 1, 2, 3, 4, 5, 6, 7, 8, 9, or more times to the recurrentparent. The parental plant that contributes the locus/loci for thedesired characteristic(s) is termed the “nonrecurrent” or “donorparent.” This terminology refers to the fact that the nonrecurrentparent is generally used one time in the breeding e.g., backcross)protocol and therefore does not recur. The locus that is transferred canbe a native gene, a mutated native gene (e.g., naturally occurring, bychemical or radiation mutagenesis, or by genome editing) or a transgeneintroduced by genetic engineering techniques into the plant (or ancestorthereof). The parental plant into which the locus/loci from thenonrecurrent parent are transferred is known as the “recurrent” parentas it is used for multiple rounds in the backcrossing protocol. Poehlman& Sleper (1994) and Fehr (1993). In a typical backcross protocol, theoriginal variety of interest (recurrent parent) is crossed to a secondvariety (nonrecurrent parent) that carries the locus/loci of interest tobe transferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until a plantis obtained wherein essentially all of the desired morphological andphysiological characteristics of the recurrent parent are recovered inthe converted plant in addition to the transferred locus/loci andassociated trait(s) from the nonrecurrent parent.

Genetic Analysis of Lettuce Cultivar Chalone.

The invention further provides a method of determining a geneticcharacteristic of lettuce cultivar Chalone or a progeny thereof, e.g., amethod of determining a genotype of lettuce cultivar Chalone or aprogeny thereof. In embodiments, the method comprises detecting in thegenome of a Chalone plant, or a progeny plant thereof, at least a firstpolymorphism (e.g., using nucleic acid amplification, nucleic acidsequencing and/or one or more molecular markers). To illustrate, inembodiments, the method comprises obtaining a sample of nucleic acidsfrom the plant and detecting at least a first polymorphism in thenucleic acid sample. Optionally, the method may comprise detecting aplurality of polymorphisms (e.g., two or more, three or more, four ormore, five or more, six or more, eight or more or ten or morepolymorphisms, etc.) in the genome of the plant. In representativeembodiments, the method further comprises storing the results of thestep of detecting the polymorphism(s) on a computer readable medium. Theinvention further provides a computer readable medium produced by such amethod.

Deposit Information

Applicants have made a deposit of at least 625 seeds of lettuce cultivarChalone with the American Type Culture Collection (ATCC), 10801University Boulevard, Manassas, Va., 20110-2209 U.S.A. under ATCCDeposit No PTA-127169 on Dec. 22, 2021. This deposit of lettuce varietyChalone will be maintained in the ATCC depository, which is a publicdepository, for a period of 30 years, or 5 years after the most recentrequest, or for the effective life of the patent, whichever is longer,and will be replaced if any of the deposited seed becomes nonviableduring that period. Additionally, Applicants have satisfied all therequirements of 37 C.F.R. §§ 1.801-1.809, including providing anindication of the viability of the samples. Access to this deposit willbe made available during the pendency of this application to theCommissioner upon request. Upon the issuance of a patent on the variety,the variety will be irrevocably and without restriction released to thepublic by providing access to the deposit of at least 625 seeds of thevariety with the ATCC. Applicants impose no restrictions on theavailability of the deposited material from the ATCC; however,Applicants have no authority to waive any restrictions imposed by law onthe transfer of biological material or its transportation in commerce.Applicants do not waive any infringement of its rights granted underthis patent or under the Plant Variety Protection Act (7 USC § 2321 etseq.).

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be apparent that certain changes and modifications suchas single locus modifications and mutations, somaclonal variants,variant individuals selected from large populations of the plants of theinstant inbred and the like may be practiced within the scope of theinvention.

What is claimed is:
 1. A seed of lettuce cultivar Chalone, arepresentative sample of seed having been deposited under ATCC AccessionNo. PTA-127169.
 2. A plant of lettuce cultivar Chalone, a representativesample of seed having been deposited under ATCC Accession No.PTA-127169.
 3. A lettuce plant, or a part thereof, having all of thephysiological and morphological characteristics of the lettuce plant ofclaim
 2. 4. A progeny lettuce plant of the plant of claim 2 thatcomprises at least 50% of the alleles of the plant of claim 2, whereinthe progeny lettuce plant comprises heavy head weight and large headcircumference.
 5. A seed that produces the plant of claim
 4. 6. A plantpart of the lettuce plant of claim
 2. 7. The plant part of claim 6,wherein the plant part is a leaf, pollen, an ovule, an anther, a root,or a cell.
 8. A tissue culture of regenerable cells of the plant ofclaim
 2. 9. A lettuce plant regenerated from the tissue culture of claim8, wherein said lettuce plant comprises all of the physiological andmorphological characteristics of lettuce cultivar Chalone.
 10. Aconverted lettuce plant produced by introducing a single locusconversion into the plant of claim 2, wherein said converted lettuceplant comprises said single locus conversion and otherwise comprises allof the physiological and morphological characteristics of lettucecultivar Chalone.
 11. A processed product from the plant of claim 2,wherein the processed product comprises cut, sliced, ground, pureed,dried, canned, jarred, washed, packaged, frozen and/or heated leaves.12. A method of producing lettuce seed, the method comprising crossingthe plant of claim 2 with itself or a second lettuce plant andharvesting the resulting seed.
 13. An F1 lettuce seed produced by themethod of claim
 12. 14. A F1 lettuce plant, or a leaf, pollen, an ovule,an anther, or a root thereof, produced by growing the seed of claim 13.15. A method for producing a seed of a lettuce plant derived from theplant of claim 2, the method comprising: (a) crossing a plant of lettucecultivar Chalone with a second lettuce plant; and (b) allowing seed toform; (c) growing a plant from the seed of step (b) to produce a plantderived from lettuce cultivar Chalone; (d) selfing the plant of step (c)or crossing it to a second lettuce plant to form additional lettuce seedderived from lettuce cultivar Chalone; and (e) optionally repeatingsteps (c) and (d) one or more times to generate further derived lettuceseed from lettuce cultivar Chalone, wherein in step (c) a plant is grownfrom the additional lettuce seed of step (d) in place of growing a plantfrom the seed of step (b).
 16. A seed produced by the method of claim15, wherein the seed comprises at least 50% of the alleles of lettucecultivar Chalone and is within one breeding cross of lettuce cultivarChalone, and wherein the seed produces a lettuce plant that comprises aheavy head weight and large head circumference.
 17. A plant, or partthereof, produced by growing the seed of claim
 15. 18. A method ofvegetatively propagating the plant of claim 2, the method comprising:(a) collecting tissue capable of being propagated from a plant oflettuce cultivar Chalone; (b) cultivating the tissue to obtainproliferated shoots; (c) rooting the proliferated shoots to obtainrooted plantlets; and (d) optionally, growing plants from the rootedplantlets.
 19. A lettuce plantlet or plant obtained by the method ofclaim 18, wherein the lettuce plantlet or plant comprises all of thephysiological and morphological characteristics of lettuce cultivarChalone.
 20. A method of introducing a desired added trait into lettucecultivar Chalone, the method comprising: (a) crossing the plant of claim2 with a lettuce plant that comprises a desired added trait to produceF1 progeny; (b) selecting an F1 progeny that comprises the desired addedtrait; (c) crossing the selected F1 progeny with lettuce cultivarChalone to produce backcross progeny; (d) selecting a backcross progenycomprising the desired added trait; and (e) optionally repeating steps(c) and (d) one or more times to produce a plant derived from lettucecultivar Chalone comprising a desired added trait and otherwiseessentially all of the physiological and morphological characteristicsof lettuce cultivar Chalone, wherein in step (c) the selected backcrossprogeny produced in step (d) is used in place of the selected F1 progenyof step (b).
 21. The method of claim 20, wherein the desired added traitis male sterility, pest resistance, insect resistance, diseaseresistance, herbicide resistance, or any combination thereof.
 22. Alettuce plant produced by the method of claim 20, wherein the lettuceplant has the desired added trait and otherwise all of the physiologicaland morphological characteristics of lettuce cultivar Chalone.
 23. Aseed of the plant of claim 22, wherein the seed produces a plant thathas the desired added trait and otherwise all of the physiological andmorphological characteristics of lettuce cultivar Chalone.
 24. A seedthat produces the plant of claim
 22. 25. A method of producing a plantof lettuce cultivar Chalone comprising a desired added trait, the methodcomprising introducing a transgene conferring the desired trait into theplant of claim 2, wherein the plant the desired added trait andotherwise comprises all of the physiological and morphologicalcharacteristics of lettuce cultivar Chalone.
 26. A lettuce plantproduced by the method of claim 25, wherein the lettuce plant comprisesthe desired added trait and otherwise all of the physiological andmorphological characteristics of lettuce cultivar Chalone.
 27. A seed ofthe plant of claim 26, wherein the seed produces a plant that has thedesired added trait and otherwise all of the physiological andmorphological characteristics of lettuce cultivar Chalone.
 28. A methodof determining a genotype of lettuce cultivar Chalone, the methodcomprising: (a) obtaining a sample of nucleic acids from the plant ofclaim 2; and (b) detecting a polymorphism in the nucleic acid sample.29. A method of producing a lettuce leaf, the method comprising: (a)growing the lettuce plant according to claim 2 to produce a lettuceleaf; and (b) harvesting the lettuce leaf.
 30. A method of producing alettuce leaf, the method comprising: (a) growing the lettuce plantaccording to claim 22 to produce a lettuce leaf; and (b) harvesting thelettuce leaf.
 31. A method of developing a lettuce line in a lettuceplant breeding program using plant breeding techniques, which includeemploying a lettuce plant, or its parts, as a source of plant breedingmaterial, the method comprising: (a) obtaining the lettuce plant, or itsparts, of claim 2 as a source of breeding material; and (b) applyingplant breeding techniques.